Inheritance of the resistance to Leptosphaeria maculans of Brassica nigra and B. juncea in near-isogenic lines of B. napus

The inheritance of resistance to Leptosphaeria maculans was studied in near-isogenic lines derived from asymmetric somatic hybrids between Brassica napus+Brassica nigra and Brassica napus+Brassica juncea, respectively. The hybrids had been backcrossed to B. napus for seven generations before the genetic segregation of the blackleg resistance was determined. The results of the inheritance studies suggested that one single dominant allele controls the resistance in the Brassica napojuncea line, whereas two independent dominant loci were found in the Brassica naponigra line. Total leaf DNA from the near-isogenic lines was isolated and 89 loci were detected by hybridization to 66 restriction fragment length polymorphism (RFLP) markers previously mapped in the B. nigra genome. Out of the 89 loci, eight loci were detected in the B. naponigra line and six were found in the B. napojuncea line. RFLP markers co-segregating with blackleg resistance in adult leaves were also found. Two markers associated with linkage group 5 and 8, respectively, of the B genome were found in the B. naponigra line and one marker was associated with linkage group 2 in the B. napojuncea line.     

Identification of a Brassica juncea-derived recessive gene conferring resistance to Leptosphaeria maculans in oilseed rape

Screening of 212 spring type Brassica napus lines carrying B genome chromosome additions and introgressions from B. nigra, B. juncea and B. carinata resulted in the identification of one line segregating for resistance to Leptosphaeria maculans (anamorph Phoma lingam) at the seedling (cotyledon) stage. This line was derived from an interspecific hybrid containing the B genome of B. juncea. Trypan blue staining of cotyledons from resistant individuals demonstrated a hypersensitive response which is delayed in plants with intermediate lesion size. Genetic analysis supported the hypothesis of a monogenic recessive inheritance of resistance. The resistance gene, termed r_jlm2, is effective in spring and winter type oilseed rape backgrounds against all tested virulent pathotypes, including two isolates which have been shown to overcome two dominant (race‐specific) B genome‐derived resistance genes in B. napus.     

Glucosinolates and Resistance to Leptosphaeria maculans in Wild and Cultivated Brassica Species

Synthetic lines of Brassica napus were derived by combining the genomes of B. atlantica and B. oleracea var. alboglabra, which were respectively resistant and susceptible to foliar infection by Leptosphaeria maculans, with a susceptible line of B. rapa. Resistance was expressed in the synthetic lines containing the genome of B. atlantica. The high levels of alkenyl glucosinolates which occur in leaves of B. atlantica, and which have been implicated in disease resistance, were also expressed within the synthetic lines, although the dominant glucosinolate had changed from sinigrin to glucobrassicanapin. Disease resistance and glucosinolate profiles did not co-segregate in F₂ progeny from crosses between the synthetic lines.     

Identification of a single dominant allele for resistance to blackleg in Brassica napus'Surpass 400'

The inheritance of resistance to blackleg (caused by Leptosphaeria maculans) was examined in the F1 and F2 of a cross between highly resistant canola ‘Surpass 400’ and susceptible ‘Westar’ in the field. Blackleg‐infected canola straw was collected from the field and scattered among plants to increase disease development with the aid of natural rainfall. Disease severity on seedlings was assessed as the average number of lesions on leaves 1 and 2, and on adult plants as the percentage necrosis on a cross‐section of stems immediately above the crown. All ‘Westar’ plants were susceptible (S) and all ‘Surpass 400’ and F1 plants were resistant (R) at both growth stages. Disease severity on F2 plants segregated 3 : 1 (R : S) as expected for a single dominant resistance allele in both the seedling and adult plant stages. There was a high proportion (91.1%) of matching reactions (R‐R and S‐S) between seedling and adult plants. ‘Surpass 400’ is the source of a single dominant allele for blackleg resistance in Brassica napus that is expressed strongly in both seedlings and adult plants.     

Different Behavior of Brassica juncea and B. carinata as Sources of Phoma lingam Resistance in Experiments of Interspecific Transfer to B. napus

With the aim to transfer Phoma lingam resistance into rape, successful interspecific crosses were made between three oilseed rape varieties (Brassica napus) and the resistant species B. carinata and B. carinata. Although both hybrid types B. napus×B. juncea and B. napus×B. carinata showed the same high level of resistance as the respective resistant parent, the resistance could be only transferred from juncea crosses. After three backcross generations, lines morphologically undistinguishable from rape, fertile, and with a high degree of resistance were obtained. The resistance of B. carinata was practically lost in the first backcross. A possible explanation of this different behavior could be a higher recombination between the genomes B and C (juncea crosses) than between B and A (carinata crosses). The: applied embryo culture increased the yield of hybrids and first backcross plants and reduced considerably the generation time.     

Haploid plantlet screening in the development of blackleg resistant DH lines of Brassica napus

This study was conducted to expedite disease-resistance breeding in canola by screening haploid plants against blackleg disease. Microspore-derived haploid plantlets from 14 unrelated crosses were inoculated with Leptosphaeria maculans pycnidiospores and were rated as resistant, intermediate or susceptible. Blackleg-inoculated and control haploid plants were then colchicine-treated to produce doubled haploid (DH) lines. The DH lines thus produced were again screened against blackleg using the cotyledon bioassay. In general, the proportion of the resistant DH lines derived from selected resistant haploid plants was much higher than the proportion of resistant DH lines from control haploid populations. There was no detrimental effect on the survival of haploid plants after the combined treatment of blackleg inoculation and colchicine. The importance of the micro-environment on the final outcome of plant-pathogen interaction in case of non-obligate pathogens is also discussed.     

Genes for race-specific resistance against blackleg disease in Brassica napus L.

Specificity of interaction at the cotyledon stage was recently demonstrated between the blackleg pathogen, Leptosphaeria maculans, and Brassica napus. Three pathogenicity groups were distinguished, PG2 avirulent towards ‘Quinta’ and ‘Glacier’, PG3 avirulent towards ‘Quinta’, and PG4 virulent on the two cultivars. The genetic control of the interactions was investigated on both the pathogen and the plant. Tetrad analysis was performed following PG3 × PG4 and PG2 × PG4 crosses.‘Quinta’ and ‘Glacier’ were crossed with the susceptible winter oilseed rape cultivar ‘Score’. The analysis of F₁, F₂ and testcross populations suggested that the incompatible interaction between ‘Quinta’ and PG3 isolates is conditioned by the presence of the dominant single resistance allele Rlml in ‘Quinta’ and the matching avirulence gene AvrLml in L. maculans. Race-specific resistance of ‘Glacier’ to PG2 isolates was conditioned by the matching gene pair Rlm2/AvrLm2. Finally, the data suggest that two avirulence genes matching two dominant loci control the ‘Quinta’-PG2 interaction. The consequences of the occurrence of race-specific resistance in B. napus are discussed with respect to future breeding for blackleg resistance.     

Introgression of Resistance to Shattering in Brassica napus from Brassica juncea through Non-Homologous Recombination

Genetic information conffering non- shattering of siliques has been introgressed in rapeseed (Brassica napus) following; interspecific hybridization between Brassica juncea and B. napus. A reconstituted B. napus plant with complete non-dehiscence of its fruits was isolated in the BC-, generation. This plant had normal meiosis and formed 19 bivalents. Its seed fertility, however, was low (23 percent). It is suggested that the gene(s) for shattering-resistance were incorporated into a B. oleracea chromosome following allosyndetic; chromosome pairing and. segmental exchange between B. nigra and B. oleracea chromosomes in the initial interspecific AABC hybrid.     

甘蓝型油菜抗菌核病研究进展

油菜是主要油料作物之一,由核盘菌(Sclerotiniasclerotiorum(Lib.)deBary)引起的菌核病(sclerotiniastemrot)是油菜的主要病害。针对当前与菌核病相关研究的新进展,本研究从4个方面对其进行了概括：（1）核盘菌的侵染方式以及在侵染过程中核盘菌分泌的草酸与寄主中钙离子的动态关系;（2）油菜通过合成植保素、酚类化合物、木质素、几丁质酶和β-1,3-葡聚糖酶等来抵抗核盘菌入侵的抗病机理;（3）现有综合防治技术中抗性种质筛选、无花瓣育种和生物防治;（4）利用与草酸代谢相关基因、抗病相关基因、防御相关转录因子基因和抗菌肽基因开展的油菜基因工程研究成果。并且进一步提出了未来油菜抗菌核病研究的可能方向,这些总结与建议为今后油菜抗菌核病研究提供了有益参考。     

Utilization of Asymmetric Somatic Hybridization for the Transfer of Disease Resistance from Brassica nigra to Brassica napus

Asymmetric somatic hybrid calli were produced between Brassica napus and a transgenic (Hyg ^R) line of B. nigra using a donor recipient fusion method for the production of cybrids. The transgenic line of B. nigra used as a donor also possessed genetic resistance to the pathogenic fungi Phoma lingam and Plasmodiophora brassicae. Using hygromycin for selection, 332 hybrid calli were obtained from which 30 produced shoots (1—-20 per callus) which were rooted on a hormone-free culture medium. The rooted shoots were transferred into soil and cultivated in a growth chamber where the plants were tested for resistance against the two pathogens. Out of 129 hybrid plants tested for resistance against P. brassicae, 30 (23.3 %) plants proved to be resistant and from 78 plants tested for resistance against P. lingam, 41 (52.6 %) plants remained disease-free after infection.     

Screening and identification of resistance to Leptosphaeria maculans (stem canker) in Brassica napus accessions

The occurrence of race-specific resistance genes to the stem canker fungus, Leptosphaeria maculans, was analysed in 453 accessions of B. napus, mainly originating from the Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK) GeneBank. Major resistance genes Rlm1, Rlm2, Rlm4 and the putative RlmBBA gene were investigated using genetically improved strains of the fungus harbouring as few corresponding avirulence genes as possible. In addition, a screening with fully virulent isolates was used to uncover novel resistance sources. Major resistance genes were rarer in frequency and diversity in spring-type cultivars compared to winter types. In the former, 65.7% of the accessions were fully susceptible to all isolates, whereas only 12.2% of the winter types were devoid of at least one R gene. In spring cultivars, the most common R gene, Rlm4 was found in 26.6% of accessions, whereas the other R genes were rare. In winter cultivars, the most common R genes were Rlm2 (more than 45.9–54.0% of the accessions) and Rlm4 (26.4–27.7% of the genotypes). In winter types however, the improvement of the quality of oils, through the generation of single- and double-low genotypes improved the homogeneity of the cvs, whereas it impoverished R gene diversity, including the loss of complete resistance that was harboured by 18.4% of the less advanced accessions, and a reduction in the ratio of accessions harbouring Rlm1. Correlation between the R gene(s) present in the accessions and their field resistance is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

甘蓝型油菜抗裂角品种(系)的选与分析

利用随机碰撞测试方法,对229份自交纯合的甘蓝型油菜品种（系）进行了抗裂角指数测定,以期筛选出抗裂角种质。结果表明,抗裂角性状在现有品种（系）中存在广泛变异,变异系数达114．4％。发现了2份抗裂角的品种（系）,占0．9％;较抗的资源占3．93％;处于中间状态的品种（系）占8．73％;易裂角资源占27．07％,极易裂角的品种（系）占59．39％。选择6个品种（系）进行了连续3年的测试,表明抗裂角性状由品种（系）的遗传特性决定,但受环境条件影响;随机碰撞法具有较好的重现性。简单相关分析显示,抗裂角指数与角果密度呈显著负相关,与角皮厚度、角果长度、角果宽度、角喙长度、角粒数呈显著正相关,但相关系数都很小。     

Brassilexin accumulation and resistance to Leptosphaeria maculans in Brassica spp. and progeny of an interspecific cross B. juncea × B. napus

Summary Resistance to Leptosphaeria maculans was assessed in Brassica napus, B. juncea, B. carinata, B. nigra and progeny issuing from an interspecific cross B. napus × B. juncea, using a cotyledon-inoculation test. In these individual plants, brassilexin accumulation was determined following an abiotic, non-specific, elicitation. All the tested B. napus cultivars were highly susceptible to the parasite and weakly accumulated brassilexin. In contrast, B. juncea, B. carinata, and B. nigra usually displayed a hypersensitive response to the inoculation and accumulated more brassilexin than B. napus. The same correlation between resistance to L. maculans and phytoalexin accumulation was observed in the interspecific hybrid progeny. The cotyledon-inoculation test allowed the discrimination of plants displaying a hypersensitive response to the inoculation from those highly sensitive to the parasite, but intermediate disease severity classes were not usually representative of resistance or susceptibility. In this respect, brassilexin determination allowed differentiation, within a set of plants presenting an intermediate response to the pathogen, of plants with a high (B. juncea-like), and with a weak (B. napus-like) ability to accumulate brassilexin.Abbreviations IHP interspecific hybrid progeny - JR B. juncea-type complete resistance to blackleg (Roy, 1984) - W&D test cotyledon-inoculation test as described by Williams & Delwiche (1979)     

Contribution of host-pathogen interactions to the expression of the blackleg disease of spring rape (Brassica napus L.) caused by Leptosphaeria maculans (Desm.) Ces. et de not

Variation in the responses of selected genotypes of spring rape to infection by a wide range of isolates of Leptosphaeria maculans was analyzed in this study.Differences in severity of disease damage to seedlings between host and pathogen genotypes were highly significant. Interactions between host and pathogen were also highly significant. The generally resistant cultivar Wesreo was susceptible to a number of isolates whereas the normally susceptible cultivars Zollern-gold and Ceska were resistant to some isolates. Overall, however, Wesreo was resistant to a far greater number of isolates than were the susceptible genotypes.Significant differences in severity of disease damage were also observed between groups of isolates from different sites. All host genotypes were resistant to a substantially greater proportion of isolates from certain sites than from others. Interactions were again detected in that the ranking of sites on the basis of the proportion of isolates causing severe damage varied between host genotypes.The significance of host-pathogen interactions in the B. napus-L. maculans relationship is discussed with special reference to current views on the nature of horizontal resistance.     

甘蓝型油菜菌核病抗性的遗传分析

利用牙签接种和花瓣接种鉴定了甘蓝型油菜DH821（抗）×DHBao604（感）组合P₁、P₂、F₁、BC₁、BC₂、F₂ 6个世代群体菌核病的抗性，通过主基因-多基因混合遗传模型分析表现，牙签接种3 d的抗性受2对加性主基因+加性-显性多基因控制，BC₁、BC₂、F₂ 群体主基因遗传率为12.49%~28.48%，多基因遗传率为21.08%~27.87%；5 d的抗性由2对加性-显性-上位性主基因控制，显性及显性互作效应明显，无多基因修饰，BC₁、BC₂、F₂群体主基因遗传率为25.19%~38.69%；3~5 d的菌丝扩展抗性由2对加性-显性-上位性主基因+加性-显性多基因控制，主基因效应明显，多基因效应不明显，BC₁、BC₂、F₂ 群体主基因遗传率为0.72%~54.06%，多基因遗传率为7.34%~46.79%。花瓣接种15 d的抗性由2对加性-显性-上位性主基因+加性-显性-上位性多基因控制，2对主基因的加性效应和显性效应为－3.6051~ －3.5225，互作效应为3.5190~3.6089，主基因遗传率为0.91%~41.79%，多基因遗传率为19.32%~73.69%。     

利用野生甘蓝抗性遗传资源获得高抗菌核病甘蓝型油菜的抗性与分子标记初步分析

茵核病是甘蓝型油菜最严重的病害之一,自身抗性低,而甘蓝野生资源中存在高抗遗传资源.以部分抗性的甘蓝型油菜中双9号和高抗性的甘蓝野生资源为亲本杂交得到F1 (ACC),F1连续用中双9号回交6代,得到育性基本恢复正常植株自交,培育成了高抗茵核病材料,命名为F6.染色体计数显示,F6有38条染色体,和甘蓝型油菜(AACC,2 n=38)一致.茵核病抗性鉴定实验结果说明F6对茵核病的抗性高于中双9号而弱于甘蓝.通过在亲本和子代间比较229个分布于甘蓝型油菜19个遗传连锁群的SSR分子标记以及16个与甘蓝抗茵核病特异性位点有关的SSR分子标记,发现甘蓝抗茵核病主要特异性区段9号连锁群已渗入甘蓝型油菜基因组中.     

Production of Brassica napus × Raphanobrassica Hybrids by Embryo Rescue: An Attempt to Introduce Shattering Resistance into B. napus

Intergeneric hybridization is one of the effective methods to broaden the genetic base of cultivated species. Raphanus sativus, a member of the subtribe Raphaninae, has very hard pods at threshing time. In an attempt to transfer shattering resistance to Brassica napus, Raphanobrassica was used as the male parent in crosses with Brassica napus. Plantlets were obtained by embryo rescue and were further multiplied in vitro by micropropagation of nodal segments. Morphology, cytology and DNA analysis confirmed the hybrid nature of these plants. They were backcrosscd with Brassica napus and the progeny was raised. Plants of BC₁ and BC₂ generations showed wide variation in morphology, chromosome number and pollen fertility. Some of the plants showed up to 95 % pollen fertility and resistance to shattering, indicating the potential for developing B. napus with resistance to shattering.     

白菜型与甘蓝型冬油菜抗寒机理差异的研究

为探究白菜型与甘蓝型冬油菜抗寒机理差异的原因。以8个抗寒性不同的冬油菜品种为材料,采用大田试验和盆栽试验相结合的方法,待油菜长至5~6片真叶时,大田试验进行植物学形态,干物质积累量的测定,盆栽试验按24℃→10℃→5℃→0℃→-5℃→-10℃各48 h依次降温处理后测定生理生化指标。结果表明,白菜型冬油菜冬前生长点洼陷,幼苗匍匐生长,干物质积累主要集中在地下部分,其中白菜型冬油菜地下部鲜质量与地下部干质量较甘蓝型冬油菜平均增加了236.1%,263.0%,说明抗寒性强的冬油菜能够在营养生长阶段将光合有机产物优先运输到地下部,建立庞大的根系,为安全越冬提供代谢能量。随着温度的变化,不同类型冬油菜的生理生化活性有较大的差异,-5℃时陇油7号SOD活性较CK增加了10.7%,0℃时陇油7号CAT、POD活性较CK分别增加了24.7%,28.6%,而0℃时白菜型冬油菜SP含量较甘蓝型冬油菜平均增加了32.3%,-10℃时白菜型冬油菜的SS含量较甘蓝型冬油菜平均增加了71.4%,-10℃甘蓝型冬油菜MDA含量较白菜型冬油菜平均增加了52.8%,这说明抗寒性强的品种在低温条件下能够保护自身免受损伤,其中CAT、POD、SP是抵抗冷害的保护性物质,SOD、SS是抵御冻害的保护性物质。白菜型冬油菜比甘蓝型冬油菜在形态学及生理水平上都具有明显的优势,形态学上的优势使其有利于抵御极端低温天气,提供维持越冬及冬后返青所需的代谢能量;生理水平上,低温胁迫后保护性酶活性、调节性物质含量增加,能够有效地保护细胞膜结构,MDA含量减少,可以缓解低温对冬油菜叶片的伤害,从而保证高越冬率,为北方白菜型与甘蓝型冬油菜抗寒性研究提供理论依据。     

油菜抗根肿病资源创新与利用的研究进展与展望

根肿病是原生动物界根肿菌门根肿菌属芸薹根肿菌( Plasmodiophora brassicae Wor.)侵染引起, 并专性危害十字花科作物的一种传染性强的土传病害。近些年来, 加拿大和我国油菜的安全生产均受到根肿病的严重威胁。本文简要介绍了根肿菌的生物学特性、分类及主要防治途径, 重点总结了在根肿病抗性资源发掘、抗病基因遗传定位和克隆以及抗病育种等方面的进展, 分析了目前我国油菜根肿病综合防控过程中存在的主要问题, 并探讨与展望了相应的对策, 旨在为油菜抗根肿病遗传育种的高效顺利开展提供技术支持, 进而保障油菜产业的可持续发展。